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Alexa Fluor™ 647 Mouse Anti-Mouse H-2Kd/H-2Dd
Alexa Fluor™ 647 Mouse Anti-Mouse H-2Kd/H-2Dd

Flow cytometric analysis of H-2Kd/H-2Dd expression on BALB/c mouse splenocytes. Splenic leucocytes from a C3H (H-2k haplotype; Left Plot) or a BALB/c (H-2d haplotype; Right Plot) mouse were stained with Alexa Fluor™ 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565357; dashed line histograms) or with Alexa Fluor™ 647 Mouse Anti-Mouse H-2Kd/H-2Dd antibody (Cat. No. 567293; solid line histograms) at 0.25 µg/test. The fluorescence histograms showing H-2Kd/H-2Dd expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Flow cytometric analysis of H-2Kd/H-2Dd expression on BALB/c mouse splenocytes. Splenic leucocytes from a C3H (H-2k haplotype; Left Plot) or a BALB/c (H-2d haplotype; Right Plot) mouse were stained with Alexa Fluor™ 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565357; dashed line histograms) or with Alexa Fluor™ 647 Mouse Anti-Mouse H-2Kd/H-2Dd antibody (Cat. No. 567293; solid line histograms) at 0.25 µg/test. The fluorescence histograms showing H-2Kd/H-2Dd expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Product Details
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BD Pharmingen™
H2-D1, H-2D; H2-K1, H-2K, H2-K
Mouse (QC Testing)
Mouse C3H, also known as C3H/He, C3H/Bi IgG2a, κ
C57BL/6 × DBA/2 (BDF1) splenocytes
Flow cytometry (Routinely Tested)
0.2 mg/ml
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
567293 Rev. 1
Antibody Details
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34-1-2S

The 34-1-2S monoclonal antibody specifically recognizes the mouse histocompatibility 2 (H-2) alloantigens, H-2Kd and H-2Dd, that are variably expressed on most nucleated cells. These MHC class I antigens are heterodimers comprised of a polymorphic alpha heavy chains (~44 kDa type I transmembrane glycoprotein encoded by the H-2 gene complex) that are noncovalently associated with invariant β2-microglobulin (~11 kDa beta light chain encoded by B2m). H-2Kd and H-2Dd are involved in the positive and negative selection of CD8+ T cells in the thymus as well as MHC-restricted antigen presentation to CD8+ αβ T cells in the periphery. H-2Kd and H-2Dd molecules can also serve a ligands for activating or inhibitory receptors, such as, those encoded by the Ly49 gene family which are variably expressed on subsets of natural killer (NK) cells and T cells. The 34-1-2S antibody cross-reacts with H-2K MHC class I alloantigens of the b, s, r, q, or p haplotypes.

567293 Rev. 1
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
567293 Rev.1
Citations & References
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Development References (6)

  1. Boudreau JE, Hsu KC. Natural Killer Cell Education and the Response to Infection and Cancer Therapy: Stay Tuned.. Trends Immunol. 2018; 39(3):222-239. (Methodology). View Reference
  2. Brennan J, Mager D, Jefferies W, Takei F. Expression of different members of the Ly-49 gene family defines distinct natural killer cell subsets and cell adhesion properties. J Exp Med. 1994; 180(6):2287-2295. (Clone-specific: Blocking). View Reference
  3. Lenz A, Heufler C, Rammensee HG, et al. Murine epidermal Langerhans cells express significant amounts of class I major histocompatibility complex antigens.. Proc Natl Acad Sci U S A. 1989; 86(19):7527-31. (Clone-specific: Flow cytometry, Immunofluorescence). View Reference
  4. Ozato K, Mayer NM, Sachs DH. Monoclonal antibodies to mouse major histocompatibility complex antigens IV. A series of hybridoma clones producing anti-H-2d antibodies and an examination of expression of H-2d antigens on the surface of these cells. Transplantation. 1982; 34(3):113-120. (Immunogen: Cytotoxicity, Radioimmunoassay). View Reference
  5. Pereira RA, Simmons A. Cell surface expression of H2 antigens on primary sensory neurons in response to acute but not latent herpes simplex virus infection in vivo.. J Virol. 1999; 73(8):6484-9. (Clone-specific: Flow cytometry). View Reference
  6. Schenkel AR, Kingry LC, Slayden RA. The ly49 gene family. A brief guide to the nomenclature, genetics, and role in intracellular infection.. Front Immunol. 2013; 4:90. (Biology). View Reference
View All (6) View Less
567293 Rev. 1

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.

Comparisons, where applicable, are made against older BD technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.